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PMID: 15756539 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Suppression of Pdx-1 perturbs proinsulin processing, insulin secretion and GLP-1 signalling in INS-1 cells.

Diabetologia ·Vol. 48 ·No. 4 ·2005-04-00 ·Pages 720-31

Wang H, Iezzi M, Theander S, Antinozzi PA, Gauthier BR, Halban PA, Wollheim CB

Abstract

Mutations in genes encoding HNF-4alpha, HNF-1alpha and IPF-1/Pdx-1 are associated with, respectively, MODY subtypes-1, -3 and -4. Impaired glucose-stimulated insulin secretion is the common primary defect of these monogenic forms of diabetes. A regulatory circuit between these three transcription factors has also been suggested. We aimed to explore how Pdx-1 regulates beta cell function and gene expression patterns. We studied two previously established INS-1 stable cell lines permitting inducible expression of, respectively, Pdx-1 and its dominant-negative mutant. We used HPLC for insulin processing, adenovirally encoded aequorin for cytosolic [Ca2+], and transient transfection of human growth hormone or patch-clamp capacitance recordings to monitor exocytosis. Induction of DN-Pdx-1 resulted in defective glucose-stimulated and K+-depolarisation-induced insulin secretion in INS-1 cells, while overexpression of Pdx-1 had no effect. We found that DN-Pdx-1 caused down-regulation of fibroblast growth factor receptor 1 (FGFR1), and consequently prohormone convertases (PC-1/3 and -2). As a result, DN-Pdx-1 severely impaired proinsulin processing. In addition, induction of Pdx-1 suppressed the expression of glucagon-like peptide 1 receptor (GLP-1R), which resulted in marked reduction of both basal and GLP-1 agonist exendin-4-stimulated cellular cAMP levels. Induction of DN-Pdx-1 did not affect glucokinase activity, glycolysis, mitochondrial metabolism or ATP generation. The K+-induced cytosolic [Ca2+] rise and Ca2+-evoked exocytosis (membrane capacitance) were not abrogated. The severely impaired proinsulin processing combined with decreased GLP-1R expression and cellular cAMP content, rather than metabolic defects or altered exocytosis, may contribute to the beta cell dysfunction induced by Pdx-1 deficiency.

MeSH Terms
Adenosine Triphosphate/metabolism Animals Calcium Signaling/physiology Cell Line, Tumor Cyclic AMP/metabolism Dose-Response Relationship, Drug Doxycycline/pharmacology Exocytosis/physiology Gene Expression/drug effects,genetics Gene Expression Regulation, Neoplastic/drug effects Glucagon-Like Peptide-1 Receptor Glucokinase/genetics Glucose/metabolism,pharmacology Glycolysis Homeodomain Proteins/genetics,metabolism,physiology Human Growth Hormone/genetics,metabolism Insulin/metabolism Insulin Secretion Islets of Langerhans/drug effects,metabolism Mitochondria/metabolism Mutation Proinsulin/metabolism Proprotein Convertases/genetics RNA, Messenger/genetics,metabolism Rats Receptor Protein-Tyrosine Kinases/genetics Receptor, Fibroblast Growth Factor, Type 1 Receptors, Fibroblast Growth Factor/genetics Receptors, Glucagon/genetics,physiology Signal Transduction/physiology Time Factors Trans-Activators/genetics,metabolism,physiology Transfection
Chemicals
GLP1R protein, human Glp1r protein, rat Glucagon-Like Peptide-1 Receptor Homeodomain Proteins Insulin RNA, Messenger Receptors, Fibroblast Growth Factor Receptors, Glucagon Trans-Activators pancreatic and duodenal homeobox 1 protein Human Growth Hormone Adenosine Triphosphate Proinsulin Cyclic AMP Glucokinase Fgfr1 protein, rat Receptor Protein-Tyrosine Kinases Receptor, Fibroblast Growth Factor, Type 1 Proprotein Convertases Glucose Doxycycline
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Wang H
Department of Cell Physiology and Metabolism, University Medical Center, 1211 Geneva 4, Switzerland. Haiyan.Wang@medicine.unige.ch
Iezzi M
Theander S
Antinozzi P A
Gauthier B R
Halban P A
Wollheim C B
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Article Info
Journal
Diabetologia
Abbr.
Diabetologia
ISSN
0012-186X
DOI
10.1007/s00125-005-1692-8
Email
haiyan.wang@medicine.unige.ch
Published
2005-04-00
Epub
2005-00-09
Pages
720-31
Language
English
Region
Germany
NLM ID
0006777
Subset
IM
External Links
PubMed source DOI: 10.1007/s00125-005-1692-8 View journal details
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